The present invention relates to a liquid processing method making use of a pipette device and an apparatus for the same, with which works including for quantifying, separating, taking out, pipetting, clarifying, condensing, diluting a liquid or a target high molecular substances included in a liquid such as useful substances such as antibiotic substance, genetic substances such as DNA, and immunological substances such as antibodies, and/or works for extracting, recovering and isolating the target high molecular substance can automatically and accurately be executed by means of absorbing and discharging the liquid through a liquid absorbing/discharging line in the pipette device.
In recent years, research activities for DNA or the like are very active in many fields including engineering, medical science, agriculture, physical science, pharmacology, and the purpose includes genome sequencing, clinical diagnosis, improvement of agricultural products, bacteriological inspection of foods, drug preparing systems or the like.
As described above, when various types of immunoassay applicable in a very wide range with high expected possibility in its application or structural analysis of molecular level organisms, microorganisms, or substances such as cells, DNA, RNA, mRNA, plasmid, virus, or bacteria (described simply as a target high molecular substance in the present specification) is performed, it is necessary to carry out with high precision works such as those for quantifying, separating, taking out, pipetting, clarifying, condensing, diluting a sample or a target high molecular substance included in the sample or works for extracting, recovering, or isolating a target high-molecular substance as a preprocessing.
To explain structural analysis of a gene such as DNA diagnosis as an example, at first it is necessary to extract, recover, and isolate a DNA region including a target gene. The technology for extracting, recovering, and isolating genes has already been established as the gene cloning technology or genome sequencing technology, and at present it is believed that, by spending enough time and expenses, any gene can be separated and obtained. For this reason, if a target gene DNA has been extracted, recovered, and separated, any type of gene analysis is possible as a principle by making use of the separated gene DNA.
However, in a case of man, for instance, a particular target gene DNA is one millionth or below of all genome DNA, and for this reason actually a quantity of DNA obtainable for testing is quite small, while a quantity of DNA and RNA not necessary for a particular experiment is quite large, which makes it difficult to execute analysis smoothly.
For the reasons as described above, to execute structural analysis of a gene such as DNA diagnosis, it is important to extract, recover, and isolate a DNA area including a target gene. Description is made hereinafter for a basic method of extracting, recovering, and isolating a DNA.
A DNA exist in a nucleus as a complex with a protein in a cell. In the basic sequence for extracting a DNA, a cell or a cell nucleus is processed with SDS (surfactant dodecil sodium sulfate) to make the DNA soluble, and proteins contained therein are removed with a proteolytic enzyme or phenol.
In other words, when a DNA is separated from the tissue, at first the tissue taken out is put in ice and kept therein for a certain period of time under a low temperature, then this cooled tissue is divided to small pieces each with the weight of around 0.1 g, which are washed with a ice-cooled buffer solution A (0.01 M Tris HCl, pH 7.8, 0.1 M NaCl, 2 mM MgCl2). This tissue is put in the above-described buffer solution A having a volume 20 times larger as compared to that of the tissue and is homogenized 5 or 10 times with a Potter type homogenizer. Then the tissue is put in a centrifugal tube together with the buffer solution and is subjected to centrifugation (2,000 rpm, for 5 minutes). The cell nucleus or the cell precipitates, so that the supernatant is aborted. When extracting a DNA from cultured cells, the cells are well suspended in an ice-cooled buffer solution B (0.01 M Tris HCl, pH 7.8, 0.1 M NaCl, 2 mM EDTA) and is subjected to centrifugation. The precipitated nuclei or cells are again well suspended in the buffer solution B having a volume 100 times larger than that of the specimen.
After the cells or cell nuclei are well suspended until a block of cells disappears, a 10% SDS solution is added by one twentieth thereof to lyse the cells. Then proteinase K (10 mg/ml) is added by one fiftieth thereof to the solution and reacted for 4 hours under a temperature of 50xc2x0 C. so that the protein is lysed. During this reaction, the solution is sometimes agitated because the viscosity is high. Then phenol extraction is executed 3 times. In this step, the extracting work should be performed carefully so that no physical power is not loaded thereto.
Then the specimen is dialyzed for around 18 hours with a buffer solution C (10 mM Tris HCl, pH 7.8/0.1 mM EDTA) having a volume 100 times larger as compared to that of the specimen, and is kept under a temperature of 4xc2x0 C.
Through the steps as described above, about 0.2 mg of DNA can be obtained from 0.1 g of tissue. What is described above is a process of extracting DNA from tissue or cells, and in addition there have been known a method in which plasmid DNA is obtained by way of the alkali method (the small quantity adjusting method), a method of recovering DNA by way of the boiling method, and a method of recovering closed-cycle bromide DNA by way of the large quantity adjusting method.
As described above, it is possible to extract, recover, and isolate DNA for structural analysis of a gene in, for instance, DNA diagnosis according to any of the known methods as described above, but a work for isolating DNA from the tissue or cells as described above is, as clearly understood from the sequence for extracting DNA from the tissue or cells as described above, extremely complicated, and a long period of time is required, which is disadvantageous.
In addition, any of various types of method including the centrifuging method, high speed liquid chromatography method, gel electrophoresis method, dispo-column method, dialysis method, glass powder method, magnetic particle cleaning nozzle method has been employed for structural analysis of DNA or the like extracted by the above-described means, and each of the methods has respective advantages and disadvantages, and at present a high precision and stable method for structural analysis has not been developed yet.
Namely, in a case of centrifugation, automation of processes for loading and taking out vessels is very difficult, and also it is very difficult to mechanically separate supernatant from precipitates after centrifugation, and for this reason its applicability for various purpose is disadvantageously poor.
In a case of high speed liquid chromatography, a separation column is basically consumable, injection for a sample to the column or time management for separation can not be mechanized, and also different samples pass through the column, which disadvantageously makes it impossible to completely prevent contamination of the column.
Furthermore, in a case of gel electrophoresis, adjustment of gel can not be mechanized, and this method has generally been used as a basic technique for separation of DNA, but the separated pieces must be taken out manually, which is disadvantageous.
The dispo-column method is one of technic which can be embodied as a kit for separation of a particular DNA piece, but the cost is very expensive, and its applicability is narrow. In addition, controls over pipetting and liquid passing through the column are difficult, and there are many problems in mechanization of this method.
In the dialysis method, a long period of time is required for dialysis, and also it is hard to apply this method when a quantity of sample is small, so that this method has not been used widely.
The glass powder method is an excellent method of extracting DNA making use of silicon dioxide, and the process is simple and convenient, but as the powder is separated with a filter or by way of centrifugation, it is difficult to automate the entire process.
Furthermore in a case of the magnetic particle cleaning nozzle method, the process can be automated by controlling the cylinder and attracting/discharging with magnetic particles, but basically it is impossible to prevent contamination only by cleaning the nozzle.
The present invention was made under the circumstances as described above, and its object is to provide a liquid processing method as well as an apparatus for the same making use of a completely novel pipette device which can automatically and with high precision execute works of quantifying, separating, taking out, pipetting, clarifying, condensing, diluting a liquid or a target high molecular substance contained in a liquid as well as works of extracting, recovering, and isolating the substance by controlling the pipette device""s operations for sucking or discharging a liquid and magnetic particles with a magnetic body and/or by a combination of a magnetic body and a filter.
Technological basis of the present invention is a liquid processing method making use of a pipette device which sucks a liquid containing a target high molecular substance via a chip detachably set in a sucking port or a discharging port of a liquid sucking/discharging line from inside of a vessel and transfers the liquid or the target high molecular substance to a target next processing position, and the chip has the sucked target high molecular substance deposited on magnetic particles and/or separated with a filter set in the chip. Namely, it is possible to automatically execute with high precision the works of quantifying, separating, taking out, pipetting, clarifying, condensing, diluting a liquid or a target high molecular substance as well as works of extracting, recovering, and isolating the substance by controlling the pipette device""s operations for sucking and discharging the liquid and magnetic particles with a magnetic body and/or by a combination of a magnetic body and a filter.
Also in the present invention, the target high molecular substance is a useful substance such as antibiotics, genetic substances such ad DNA, or an immunological substance such as antibody. For this reason, the present invention is well suited to works of separating, taking out, pipetting, clarifying, condensing, diluting and/or works of capturing, extracting, isolating, amplifying, labelling, and measuring molecule level organisms or microorganisms such as cells, DNA, RNA, mRNA, plasmid, virus, and bacteria or certain high molecular substance, and a target high molecular substance can be obtained without depending on the conventional centrifugation.
Also in the present invention, such works as quantifying, separating, taking out, pipetting, cleaning, condensing, and diluting the target high molecular substance as described above is carried out with a chip set in the liquid sucking/discharging line described above and at least one type of filter set in a tip section of the chip. With this configuration, such works as quantifying, separating, taking out, pipetting, clarifying, condensing, and diluting the target high molecular substance can easily be executed with high precision.
The present invention is embodied mainly as described above, but by providing a plurality of filter holders in multiple stages in a way where, for instance, a filter holder with a filter to screen out blood corpuscle shells is provided in the first stage of the chip and a filter holder with a silica membrane filter to capture DNA is provided in the second stage thereof, such works as quantifying, separating, taking out, pipetting, clarifying, condensing, and diluting the target high molecular substance as described above can more easily be carried out with high precision. It is needless to say that, in a case where separation of a target high molecular substance is executed by setting each filter holder according to the present invention, the chips and filter holders may be engaged in and processed one by one or complied in a multiple stages and set to execute a plurality of works simultaneously.
Also in the present invention, by using a plurality of filters each having a different pore size (transmission diameter of each filter) and used for separation of a target high molecular substance and foreign materials other than the target high molecular substance respectively, it is possible to obtain only the target high molecular substance without fail.
Also in the present invention, after such works as quantifying, separating, taking out, pipetting, clarifying, condensing, diluting a liquid or a target high molecular substance with the filter as described above, in a step of detachably setting a new chip in a tip section of the liquid sucking/discharging line and sucking/discharging a solution containing magnetic particles with this chip, as the magnetic particles are attracted by a magnetic body provided in the side of the chip onto an internal surface of the chip to extract, recover, and isolate the target high molecular substance, such works as quantifying, separating, taking out, pipetting, clarifying, condensing, and diluting and also such works as extracting, recovering, and isolating a target high molecular substance can automatically be executed.
Also in the present invention, different from a liquid processing based on the filter system as described above, such works as capturing, extracting, isolating, amplifying, labelling, and measuring the target high molecular substance may be executed only with a chip set in the liquid sucking/discharging line, a magnetic force, and one or a plurality types of magnetic particles and without using the filter as described above, more precise liquid processing can be realized with simpler configuration.
Also in the present invention, by causing the chip set in the liquid sucking/discharging line as described above to react with magnetic particles like in the invention described above, refining process such as capturing cells, having cell cores or protein lysed can automatically be executed, and a particular target high molecular substance can easily be extracted, recovered, and isolated.
Furthermore in the present invention, by using magnetic particles with a probe or biotin or streptoavidin coated thereon by making use of a chip set in a liquid sucking/discharging line like in the invention described above after the work of extracting the substance, a particular base sequence piece can easily be isolated with high precision without executing centrifugation.
Also in the present invention, it is possible to execute a series of works for refining such as capturing cells, or having cell core or protein lysed by causing a chip set in the liquid sucking/discharging line to react with magnetic particles for extracting a particular high molecular substance and then isolating the particular base sequence piece with other type of magnetic particles with a probe or biotin or streptoavidin coated thereon easily in a liquid sucking/discharging line in a pipette device.
Also in the present invention, after a series of works such as capturing, extracting, and isolating a target high molecular substance by using the magnetic particles as described above, by making the isolated particular base sequence piece emit light through chemical luminescence or fluorescence or enzymatic coloration, presence or a quantity of the particular base sequence piece can easily be detected or measured.
Also in the present invention, it is possible to easily and automatically execute a series of works for refining such as capturing cells, or having cell core or protein lysed by causing a chip set in the liquid sucking/discharging line to react with magnetic particles for extracting a particular high molecular substance, then amplifying the extracted target high molecular substance, isolating the particular base sequence piece with other type of magnetic particles with a probe or biotin or streptoavidin coated thereon and then detecting presence of or measuring a quantity of the particular base sequence piece by causing the isolated piece to emit light through chemical luminescence or fluorescence or enzymatic coloration.
In the present invention, by executing the works of separating, taking out, pipetting, clarifying, condensing, diluting the target high molecular substance and/or works for capturing, extracting, isolating, amplifying, labelling, and measuring the substance in a single liquid sucking/discharging line or a plurality of liquid sucking/discharging lines provided in parallel to each other, a sequences of works can be executed efficiently and automatically. In a case where a plurality of liquid sucking/discharging lines are provided in parallel, the processing capacity is improved, and also a multi-channel processing line can be realized.
In the present invention, in a case where processing is executed with a plurality of liquid sucking/discharging lines provided in parallel, as the plurality of liquid sucking/discharging lines are driven and controlled so that the works of separating, taking out, pipetting, clarifying, condensing, diluting the target high molecular substance and/or the works of capturing, extracting, isolating, amplifying, labelling, and measuring the substance in each line are executed according to the same timing, or also so that each liquid is sucked or discharged in a specified processing step according to a different timing, and for this reason processing steps suited to a target high molecular substance can easily be built up.
In the present invention, by providing working spaces separated from each other with partitions in the single or a plurality of liquid sucking/discharging lines, or by providing working spaces with an air flow by means of continuously sucking air in each line working space from an air sucking port, or by combining these different types of configuration, even in a case of liquid processing such as extracting and analyzing DNA or the like in which it is required to strictly prevent contamination by air in each processing line, the objective can easily be achieved.
Also in the present invention, by having a target high molecular substance or a substance bonded to a target high molecular substance absorbed or bonded to a surface of each magnetic particle used for the purpose of the present invention, the target high molecular substance can be obtained without executing centrifugation.
In the present invention, in a case where the above-described magnetic particles are used, controls are provided so that the magnetic particles are absorbed onto an internal wall of a chip due to a magnetic force working from outside of the chip, or so that, if effect of the magnetic force is weak or not present, the magnetic particles are held separable from the internal surface of the chip, it is possible to control capture of target high molecular substance and separation of the same from foreign materials with high precision.
In the present invention, by controlling load of a magnetic force into or elimination of a magnetic force in a chip by means of moving a permanent magnet in a direction perpendicular to the longitudinal direction of the chip or in a range including the direction perpendicular to the longitudinal direction of the chip, or by turning ON or OFF an electric magnet, it is possible to efficiently execute such works as absorbing magnetic particles or agitating and mixing the magnetic particles with other liquid or cleaning.
In the present invention, in a case where processing is executed with an electric magnet, by turning ON the electric magnet when it contacts an external surface of the chip to generate a magnetic force or by moving off the electric magnet from the chip when the magnetic force is eliminated, absorption of magnetic particles or agitation and mixing of magnetic particles with other solution or cleaning can efficiently be executed.
In the present invention, when removing a chip from a liquid sucking/discharging line, it can easily be removed by holding the chip with a holding body synchronously operating when the permanent magnet or electric magnet moves to the chip and the permanent magnet or electric magnet and then moving the liquid sucking/discharging line upward.
In the present invention, the chip described above comprises a small diameter section steeped into a liquid, a large diameter section having a capacity larger than a capacity of a vessel in which a liquid is accommodated, and an intermediate section provided between the small diameter section and the large diameter section and having a diameter smaller at least than that of the large diameter section, and magnetic particles are captured by the intermediate section, so that clogging never occurs and the magnetic particles can almost completely be absorbed because of magnetism within a short period of time.
In the present invention, an internal diameter of the intermediate section of a chip described above has a dimension appropriate for ferromagnetic field of the magnetic body to provide effects therein, and magnetic particles are captured due to a magnetic force generated in the ferromagnetic field of the magnetic body, so that magnetic particles can almost completely be captured because of magnetism within a short period of time.
In the present invention, by forming an internal diameter of the intermediate section of a chip described above so that it has the substantially same width dimension as that of a contacting surface of a magnetic body contacting the intermediate section, the most effective absorption of magnetic particles can be realized.
In the present invention, absorption of magnetic particles onto an internal surface of a chip set in the liquid sucking/discharging line is executed by passing a solution containing magnetic particles through a ferromagnetic field inside a chip at a slow speed appropriate for the magnetic particles to be captured completely, the magnetic particles can completely be captured.
In the present invention, controls are provided so that the final liquid surface of a liquid passing through the chip when sucked into or discharged from the chip always reaches the magnetic field described above, so that the magnetic particles can be captured more completely.
In the present invention, in a case where a liquid is sucked or discharged with a so-called single nozzle, the liquid is sucked by contacting a tip section for a chip set in the liquid sucking/discharging line to an internal bottom of a vessel with the liquid accommodated therein and then lightly raising the tip section, so that almost all of the liquid within the vessel can be sucked and uniformity of reaction can be maintained.
In the present invention, agitation and mixing of magnetic particles absorbed in the chip with a reagent or cleaning water is executed under the so-called pumping control in which the works of sucking and discharging the liquid in the liquid sucking/discharging line is continuously executed at a high speed and times enough to agitate and mix the liquid with the magnetic particles, so that the liquid and magnetic particles can homogeneously be agitated and mixed with each other.
In the present invention, in a case where the works of agitating and mixing a liquid and magnetic particles with each other are executed, the works of sucking and discharging the liquid in the liquid sucking/discharging line is executed with the tip section steeped into a reagent or cleaning water accommodated in a vessel so that a quantity of a liquid in the vessel and a sucking/discharging rate substantially coincide with each other, and for this reason no bubble is generated and the reaction can be executed under no physical impact, so that separation of a target high molecular substance from the magnetic particles due to bubbles can be prevented without fail.
In the present invention, in a case where necessary temperature controls are provided to promote a reaction between a target high molecular substance and a reagent or the like or amplification of the target high molecular substance, the reaction liquid or a liquid to be amplified is transferred with the chip into each thermostatic chamber previously kept in a specified temperature to be heated or cooled therein, so that a period of time required for heating or cooling the reaction liquid or the liquid to be amplified can substantially be reduced.
In the present invention, when controlling a temperature in each reaction chamber, a covering body is set over a tip section of the liquid sucking/discharging line, and the covering body is set via the liquid sucking/discharging line on the thermostatic chamber, so that evaporation of the liquid can be prevented and also contamination of air can be prevented without fail.
In the present invention, the covering body is built so that it is plucked and broken by the liquid sucking/discharging line or a chip set in the line, and for this reason it is not necessary to provide a separate sucking/discharging means, and a reaction liquid or a liquid to be amplified in a thermostatic vessel can be sucked by the liquid sucking/discharging line or a chip set therein, so that the configuration is quite simple and a series of works can automatically be executed.
In the present invention, to realize each of the liquid processing methods as described above, there is provided a liquid processing apparatus making use of a pipette device comprising a liquid sucking/discharging line which can move in the horizontal direction and is maintained at a specified position so that it can move in the vertical direction, a means for executing liquid sucking/discharging works through the liquid sucking/discharging line, a plurality of chips required for processing one type of liquid and provided along the horizontal direction in which the liquid sucking/discharging line moves, a vessel with the liquid accommodated therein, one or more filter holders each having a filter required for the processing above, and one or more vessels each having other type of liquid required for the processing above, and the liquid sucking/discharging line or a chip set therein is driven and controlled according to instructions from a control unit so that the chip is transferred with a filter holder set therein to execute such works as quantifying, separating, taking out, pipetting, clarifying, condensing, and diluting the liquid or a target high molecular substance contained in the liquid, and for this reason it is not necessary to especially provide a means in which an operation may be interrupted such as a centrifugal separator, and it is possible to automate such works as quantifying, separating, taking out, pipetting, clarifying, condensing, diluting a target high molecular substance with simple configuration.
In the present invention, there is provided a liquid processing apparatus comprising a liquid sucking/discharging line which can move in the horizontal direction and is maintained at a specified position so that it can move in the vertical direction, a means for executing liquid sucking/discharging works through the liquid sucking/discharging line, a plurality of chips required for processing one type of liquid and provided along the horizontal direction in which the liquid sucking/discharging line moves, a vessel with the liquid accommodated therein, a magnetic body for attracting magnetic particles contained in a liquid onto an internal surface of a chip when the liquid is sucked into or discharged from the chip, one or more vessels with other types of liquid accommodated therein respectively required for the processing described above, and the liquid sucking/discharging line or the chip is driven and controlled according to instructions from a control unit so that the chip is transferred to execute such works as capturing, extracting, isolating, amplifying, labelling, and measuring a liquid or a target high molecular substance contained in the liquid, and for this reason it is not necessary to especially provide a means in which the operation such as a centrifugal separator may be interrupted, and also it is possible to automate such works as capturing, extracting, isolating, amplifying, labelling, and measuring a target high molecular substance with simple configuration.
In the present invention, there is provided a liquid processing apparatus making use of a pipette device comprising a liquid sucking/discharging line which can move in the horizontal line and is maintained at a specified position so that it can move in the vertical direction, a plurality of chips required for processing one type of liquid and provided along the horizontal line in which this sucking/discharging line moves, a vessel with the liquid accommodated therein, one or more filter holders each having a filter set therein required for the processing described above, one or more vessels each accommodating therein other types of liquid required for the processing above, a vessel in which a liquid containing magnetic particles is accommodated, and a magnetic body for attracting the magnetic particles onto an internal surface of the chip in the process of sucking or discharging a solution containing the magnetic particles, and the liquid sucking/discharging line is transferred according to instructions from a control unit to execute required processing for a liquid or a target high molecular substance contained in the liquid, and for this reason it is possible to execute such works as quantifying, separating, taking out, pipetting, clarifying, condensing, diluting a target high molecular substance and also such complicated works as extracting, recovering, and isolating the target high molecular substance with very simple configuration in succession and automatically.
In the present invention, a hook for locking and supporting a chip engaged in and supported by the liquid sucking/discharging line is rotatably supported by the liquid sucking/discharging line, and the hook is energized in its normal state in the direction in which connection between the liquid sucking/discharging line and the chip is maintained, and also the hook is energized by a lock releasing body provided at a specified position in the direction in which locking between the liquid sucking/discharging line and the chip is released, so that it is possible, when a filter holder is set in or removed from a tip section of a chip, to prevent the chip from being separated from the liquid sucking/discharging line without fail, and also locking with the hook can automatically be released.
In the present invention, in a case of a liquid processing apparatus with the hook attached thereto, the filter holder set in a tip section of the chip is transferred so that the chip and/or the filter holder is separated from an edge of the liquid sucking/discharging line or a chip set therein when the liquid sucking/discharging line locked by the locking body is moved upward, so that the work of removing a chip and/or a filter holder can be automated.
In the present invention, the vessel used according to the present invention is formed into a cassette form having a plurality of chambers each for accommodating a type of liquid therein and then samples or reagents required for a reaction or processing can be pipetted to each of the liquid accommodating section, so that high precision liquid processing can be realized. In this case, a portion or all of the reagent previously accommodated in each liquid accommodating section is shielded with a thin film body which can be broken by the liquid sucking/discharging line or a chip set thereon, and a mechanism for pipetting each reagent becomes unnecessary, which is desirable for simplifying configuration of an apparatus.
In the present invention, in a case where the magnetic body is built with a permanent magnet, a surface of the permanent magnet contacting a chip is formed according to an external form of the chip and the chip is movably provided in a direction perpendicular to the longitudinal direction of the chip, so that it is possible not only to completely capture magnetic particles, but also to prevent adverse effects by diffusion and movement of the magnetic particles in association with the magnet without fail.
In the invention, the magnetic body is built with an electric magnet in place of the permanent magnet described above with a surface thereof contacting a chip formed according to an external form of the chip, and is provided so that the electric magnet generates a magnetic force when it contacts outside of the chip and also can move, when degaussed, in a direction perpendicular to the longitudinal center line of the chip or in a range including the direction, and for this reason magnetic particles are attracted in association with movement of the magnetic body along the longitudinal center line of the chip so that it is possible to prevent the magnetic particles from going out of control and control over the magnetic particles from being lost, which makes it possible to realize complete attraction of the magnetic particles.
In the present invention, a holding body, which moves, when the permanent magnet or electric magnet moves to the chip, in synchronism to movement of the magnet, is provided, a surface of the holding body contacting a chip is formed according to an external form of the chip, and the chip is held between the holding body and the permanent magnet or electric magnet, so that the chip can easily be removed only by moving the liquid sucking/discharging line upward.
In the present invention, a temperature control step required for a reaction between a target high molecular substance and a reagent or the like or for amplifying the target high molecular substance is inserted into the liquid processing step with the liquid sucking/discharging line, the reaction liquid or the liquid to be amplified is transferred with the chip to each thermostatic vessel kept at a prespecified temperature, and also a covering body, which can be set in a tip section of the liquid sucking/discharging line, is set by the liquid sucking/discharging line on each thermostatic vessel in which the reaction liquid or the liquid to be amplified is accommodated, so that also amplification of the target high molecular substance can successively be processed in a series of works.
Furthermore in the present invention, the covering body comprises a flat surface section having a diameter larger than that of a bore of the thermostatic vessel and a maintenance groove section formed in a substantially central portion of the flat surface section and having the same bore as an external diameter of the liquid sucking/discharging line or the tip of the chip, and a bottom section of the maintenance groove section is formed with a thin film body which can be broken by the liquid sucking/discharging line or the chip, so that it is not necessary to separately provide a covering body supply means or a liquid sucking/discharging means, which largely simplifies this type of apparatus.